Chicken Lines Research Articles

Highly elevated base excision repair pathway in primordial germ cells causes low base editing activity in chickens.

Base editing technology enables the generation of precisely genome-modified animal models. In this study, we applied base editing to chicken, an important livestock animal in the fields of agriculture, nutrition, and research through primordial germ cell (PGC)-mediated germline transmission. Using this approach, we successfully produced two genome-modified chicken lines harboring mutations in the genes encoding ovotransferrin (TF) and myostatin (MSTN); however, only 55.5% and 35.7% of genome-modified chickens had the desired base substitutions in TF and MSTN, respectively. To explain the low base-editing activity, we performed molecular analysis to compare DNA repair pathways between PGCs and the chicken fibroblast cell line DF-1. The results revealed that base excision repair (BER)-related genes were significantly elevated in PGCs relative to DF-1 cells. Subsequent functional studies confirmed that the editing activity could be regulated by modulating the expression of uracil N-glycosylase (UNG), an upstream gene of the BER pathway. Collectively, our findings indicate that the distinct DNA repair property of chicken PGCs causes low editing activity during genome modification, however, modulation of BER functions could promote the production of genome-modified organisms with the desired genotypes.

Novel Avian Pathogenic Escherichia coli Genes Responsible for Adhesion to Chicken and Human Cell Lines.

Avian pathogenic Escherichia coli (APEC) is a major bacterial pathogen of commercial poultry contributing to extensive economic losses and contamination of the food chain. One of the initial steps in bacterial infection and successful colonization of the host is adhesion to the host cells. A random transposon mutant library (n = 1,300) of APEC IMT 5155 was screened phenotypically for adhesion to chicken (CHIC-8E11) and human (LoVo) intestinal epithelial cell lines. The detection and quantification of adherent bacteria were performed by a modified APEC-specific antibody staining assay using fluorescence microscopy coupled to automated VideoScan technology. Eleven mutants were found to have significantly altered adhesion to the cell lines examined. Mutated genes in these 11 "adhesion-altered mutants" were identified by arbitrary PCR and DNA sequencing. The genes were amplified from wild-type APEC IMT 5155, cloned, and transformed into the respective adhesion-altered mutants, and complementation was determined in adhesion assays. Here, we report contributions of the fdtA, rluD, yjhB, ecpR, and fdeC genes of APEC in adhesion to chicken and human intestinal cell lines. Identification of the roles of these genes in APEC pathogenesis will contribute to prevention and control of APEC infections.IMPORTANCE Avian pathogenic E. coli is not only pathogenic for commercial poultry but can also cause foodborne infections in humans utilizing the same attachment and virulence mechanisms. Our aim was to identify genes of avian pathogenic E. coli involved in adhesion to chicken and human cells in order to understand the colonization and pathogenesis of these bacteria. In contrast to the recent studies based on genotypic and bioinformatics data, we have used a combination of phenotypic and genotypic approaches for identification of novel genes contributing to adhesion in chicken and human cell lines. Identification of adhesion factors remains important, as antibodies elicited against such factors have shown potential to block colonization and ultimately prevent disease as prophylactic vaccines. Therefore, the data will augment the understanding of disease pathogenesis and ultimately in designing strategies against the infections.

Molecular identification and characterisation of a novel chicken leukocyte immunoglobulin-like receptor A5

ABSTRACT 1. Leukocyte immunoglobulin-like receptor A5 (LILRA5) is a key molecule that regulates the immune system. However, the LILRA5 gene has not been characterised in avian species, including chickens. The present study aimed to identify and functionally characterise LILRA5 identified from two genetically disparate chicken lines, viz., Marek’s disease (MD)-resistant (R) line 6.3 and MD-susceptible (S) line 7.2. 2. Multiple sequence alignment and phylogenetic analyses confirmed that the identity and similarity homologies of amino acids of LILRA5 in chicken lines 6.3 and 7.2 ranged between 93% and 93.7%, whereas those between chicken and mammals ranged between 20.9% and 43.7% and 21.1% to 43.9%, respectively. The newly cloned LILRA5 from chicken lines 6.3 and 7.2 revealed high conservation and a close relationship with other known mammalian LILRA5 proteins. 3. The results indicated that LILRA5 from chicken lines 6.3 and 7.2 was associated with phosphorylation of Src kinases and protein tyrosine phosphatase non-receptor type 11 (SHP2), which play a regulatory role in immune functions. Moreover, the results demonstrated that LILRA5 in these lines was associated with the activation of major histocompatibility complex (MHC) class I and β2-microglobulin and induced the expression of the transporter associated with antigen processing. In addition, LILRA5 in both chicken lines activated and induced Janus kinase (JAK)–signal transducer and the activator of transcription (STAT), nuclear factor kappa B (NF-κB), phosphoinositide-3-kinase (PI3K)/protein kinase B (AKT) and the extracellular signal-regulated kinase (ERK)1/2 signalling pathways; toll-like receptors; and Th1-, Th2-, and Th17- cytokines. 4. The data suggested that LILRA5 has innate immune receptors essential for macrophage immune response and provide novel insights into the regulation of immunity and immunopathology.

Functional genes polymorphism associations with egg quality traits in the populations of dual-purpose

Aim. To study the egg quality traits of Poltava Clay chicken line 14 and Rhode-Island Red chicken line 38 with different genotypes of the prolactin gene (PRL), growth hormone gene (GH), growth hormone receptor gene (GHR), insulin-like growth factor I gene (IGF-I) and Mx gene (Mx). Methods. The study was conducted using the method of polymerase chain reaction and restriction fragment length polymorphism analysis (PCR-RFLP). Results. We found signifi cant differences in line 14 for egg quality between prolactin, growth hormone, growth hormone receptor and Mx loci. Homozygous individuals CC and TT by prolactin locus prevailed over heterozygotes CT for egg weight on the 30th week of life. As for the growth hormone gene, the maximum differences for egg weight were revealed when comparing BC heterozygotes with CC homozygotes. As for the growth hormone receptor gene, signifi cant prevalence (p < 0.05) of individuals with the B0 genotype over A0 by parameters of egg yolk weight was noted at the age of 52 weeks. Signifi cant differences (p < 0.05) in eggshell thickness were determined for genotypes AG and GG by Mx gene in week 52. There were signifi cant differences (p < 0.05) in egg quality traits for prolactin and Mx gene for chickens of line 38. TT homozygotes by prolactin locus are characterized by the prevalence of values (p < 0.05) for the egg, yolk and shell weight. In case of Mx gene polymorphism, the heterozygous individuals were characterized by signifi cantly higher values (p < 0.05) of egg and albumen weight on the 30th week of life. There were no signifi cant differences in both experimental chicken lines for other egg quality traits between individuals with different genotypes. Conclusions. The data obtained are recommended for the use in breeding programs for Poltava Clay chicken line 14 and Rhode-Island Red chicken line 38 with the aim of obtaining microlines with the different genotypes for PRL, GH, GHR and Mx loci.

Integrated Transcriptome and Histone Modification Analysis Reveals NDV Infection Under Heat Stress Affects Bursa Development and Proliferation in Susceptible Chicken Line.

Two environmental factors, Newcastle disease and heat stress, are concurrently negatively impacting poultry worldwide and warrant greater attention into developing genetic resistance within chickens. Using two genetically distinct and highly inbred layer lines, Fayoumi and Leghorn, we explored how different genetic backgrounds affect the bursal response to a treatment of simultaneous Newcastle disease virus (NDV) infection at 6 days postinfection (dpi) while under chronic heat stress. The bursa is a primary lymphoid organ within birds and is crucial for the development of B cells. We performed RNA-seq and ChIP-seq targeting histone modifications on bursa tissue. Differential gene expression revealed that Leghorn, compared to Fayoumi, had significant down-regulation in genes involved in cell proliferation, cell cycle, and cell division. Interestingly, we also found greater differences in histone modification levels in response to treatment in Leghorns than Fayoumis, and biological processes enriched in associated target genes of H3K27ac and H3K4me1 were similarly associated with cell cycle and receptor signaling of lymphocytes. Lastly, we found candidate variants between the two genetic lines within exons of differentially expressed genes and regulatory elements with differential histone modification enrichment between the lines, which provides a strong foundation for understanding the effects of genetic variation on NDV resistance under heat stress. This study provides further understanding of the cellular mechanisms affected by NDV infection under heat stress in chicken bursa and identified potential genes and regulatory regions that may be targets for developing genetic resistance within chickens.

Liver Transcriptome Responses to Heat Stress and Newcastle Disease Virus Infection in Genetically Distinct Chicken Inbred Lines.

Heat stress results in reduced productivity, anorexia, and mortality in chickens. The objective of the study was to identify genes and signal pathways associated with heat stress and Newcastle disease virus (NDV) infection in the liver of chickens through RNA-seq analysis, using two highly inbred chicken lines (Leghorn and Fayoumi). All birds were held in the same environment until 14 days of age. On day 14, half the birds were exposed to 38 °C with 50% relative humidity for 4 h, then 35 °C until the end of the experiment. The remaining birds were kept at 25 °C throughout the experiment. The heat-treated birds were inoculated at 21 days of age with 107 EID50 (One EID50 unit is the amount of virus that will infect 50 percent of inoculated embryos) NDV La Sota strain to investigate the effects of both heat stress and NDV infection. Physiological parameters were recorded as blood phenotypes at three stages: acute heat (AH), chronic heat (CH1), and chronic heat combined with NDV infection (CH&NDV), at 4 h, 7 days, and 10 days post-initiation of heat treatment, respectively. Our previous work revealed that the heat-resilient Fayoumi line maintained a more stable acid-base balance in their blood compared to the Leghorn line. Liver samples were harvested on both AH and CH&NDV to characterize the transcriptome profiles of these two inbred lines. Both genetic lines and treatments had large impact on the liver transcriptome. Fayoumi birds had more differentially expressed genes (DEGs) than Leghorn birds for both treatments. Metabolic and immune-related genes were on the DEG list, with Fayoumi having more immune-related DEGs than Leghorns, which was confirmed by gene functional enrichment analysis. Weighted correlation network analysis (WGCNA) indicated that the driver genes such as Solute Carrier Family genes could be very important for stabilizing the acid-base balance in Fayoumi birds during heat stress. Therefore, candidate genes such solute carrier family genes could be potential genetic targets that are regulated by Fayoumis to maintain physical hemostasis under heat stress. Differential gene expression showed that Leghorns mainly performed metabolic regulation in response to heat stress and NDV infection, while Fayoumis regulated both immune and metabolic functions. This study provides novel insights and enhances our understandings of liver response to heat stress of heat resilient and susceptible inbred chicken lines.

Genetic Variability of Indels in the Prolactin and Dopamine Receptor D2 Genes and Their Association with the Yield of Allanto-Amniotic Fluid in Russian White Laying Hens

Currently, there is virtually no information on genetic factors affecting the yield of allanto-amniotic fluid, which is the raw material for the production of human and animal vaccines. Association studies including this trait are beneficial for increasing productivity of a biotechnological line of chickens used for the production of ‘Clean Eggs’. We examined here a population of the Russian White breed for the effects of indels in the prolactin (PRL) and dopamine receptor D2 (DRD2) genes on the yield of extraembryonic fluid (YEF) and embryo weight at 12.5 days of development. A 24-bp insertion in the 5' flanking region of the PRL gene significantly (P<0.01) increases YEF in the embryos. The heterozygous embryos contained the highest YEF (9.6 mL) than that of the homozygous insertion (9.4 mL) and deletion embryos (8.4 mL). We also found a significant association (P<0.001) between the PRL genotypes and egg weight (EW). The results of the present study suggest a significant association between the PRL gene variation and quantitative traits in the Russian White chickens, contributing to a long-term programme on the effective use of the genetic potential of Russian gene pool breeds and populations of chickens.

Genetic Dissection of Growth Traits in a Unique Chicken Advanced Intercross Line.

The advanced intercross line (AIL) that is created by successive generations of pseudo-random mating after the F2 generation is a valuable resource, especially in agricultural livestock and poultry species, because it improves the precision of quantitative trait loci (QTL) mapping compared with traditional association populations by introducing more recombination events. The growth traits of broilers have significant economic value in the chicken industry, and many QTLs affecting growth traits have been identified, especially on chromosomes 1, 4, and 27, albeit with large confidence intervals that potentially contain dozens of genes. To promote a better understanding of the underlying genetic architecture of growth trait differences, specifically body weight and bone development, in this study, we report a nine-generation AIL derived from two divergent outbred lines: High Quality chicken Line A (HQLA) and Huiyang Bearded (HB) chicken. We evaluate the genetic architecture of the F0, F2, F8, and F9 generations of AIL and demonstrate that the population of the F9 generation sufficiently randomized the founder genomes and has the characteristics of rapid linkage disequilibrium decay, limited allele frequency decline, and abundant nucleotide diversity. This AIL yielded a much narrower QTL than the F2 generations, especially the QTL on chromosome 27, which was reduced to 120 Kb. An ancestral haplotype association analysis showed that most of the dominant haplotypes are inherited from HQLA but with fluctuation of the effects between them. We highlight the important role of four candidate genes (PHOSPHO1, IGF2BP1, ZNF652, and GIP) in bone growth. We also retrieved a missing QTL from AIL on chromosome 4 by identifying the founder selection signatures, which are explained by the loss of association power that results from rare alleles. Our study provides a reasonable resource for detecting quantitative trait genes and tracking ancestor history and will facilitate our understanding of the genetic mechanisms underlying chicken bone growth.

Multiphasic poultry growth models: method and application

Growth and development are complex phenomena. To date, most growth modeling research has focused on a single growth phase, which is sufficient and useful for describing ad libitum fed animals processed at a prepubertal age, such as broilers or turkeys produced for meat. However, multiphase growth models are necessary to describe and predict growth and further to hypothesize about optimizing growth of reproducing animals such as broiler breeder hens. Therefore, the objective of the present study was to develop and evaluate multiphasic models to describe the growth of various types of poultry raised to reproductive age. Coefficients for monophasic, diphasic, and triphasic Gompertz model forms were estimated using a variety of BW trajectories published by primary breeders. The fit of these models was evaluated for a representative laying line hen, broiler breeder hen and rooster, and turkey hen. The coefficient of determination (R2), root mean square error, and the Bayesian information criterion were used to evaluate the fit of each model. The diphasic model was found to be the best fit for the turkey hen, while the triphasic model was the most suitable model for all the chicken lines studied. Hypotheses can be formulated based on any of the continuous model parameters, and the resulting BW trajectories can be implemented and evaluated in a systematic way. The biological relevance of the continuous parameters in multiphasic Gompertz models provides an opportunity to implement a robust hypothesis-based approach for future optimization of growth curves.

Effect of in ovo application of hydroxyapatite nanoparticles on chicken embryo development, oxidative status and bone characteristics

ABSTRACT Intensive selection in modern lines of fast-growing chickens has caused several skeletal disorders. Therefore, current research is focused on methods to improve the bones of birds. A new potential solution is in ovo technology using nanoparticles with a high specificity for the bone tissue. Thus, the objective of the present study was to evaluate the effect of in ovo application of hydroxyapatite nanoparticles (HA-NP) in different concentrations (50, 100 and 500 μg/ml colloids) on chicken embryo development, with a particular focus on the oxidative status and bone characteristics of the embryo. The results showed that in ovo treatment with HA-NP did not negatively affect hatchability and body weight. However, bone weight was reduced in 500 μg/ml group. The concentrations of calcium, phosphorus and crude ash were not affected. The modulatory effect of HA-NP was observed on the basis of antioxidative markers – superoxide dismutase, total antioxidant status, malondialdehyde in serum and selected tissues. Glutathione concentration in serum suggested higher metabolic stress. Among bone turnover markers, the concentration of osteocalcin was found to be significantly affected by HA-NP injection. Thus, the in ovo application of HA-NP could modify the molecular responses at the stage of embryogenesis but these changes were not reflected in embryo growth and even slowed down bone development. Nevertheless, the question for the follow-up research is whether in ovo administration of HA-NP would affect the antioxidative status and bone turnover resulting in improved bone conditions and body gain in post hatch chickens.

Evaluation of Vanaraja female line chicken for growth, production, carcass and egg quality traits

Vanaraja female line (PD-2) chicken was evaluated for growth, production, carcass and egg quality traits utilizing the data generated from 3,737 chicks and 599 hens produced in pedigreed mating with 50 sires and 250 dams at ICAR-Directorate of Poultry Research, Hyderabad, Telangana. Fertility was 86.63% and the hatchability was 86.72% on fertile egg set and 72.53% on total egg set basis in PD-2 line. The least squares means (LSM) for six week body weight and shank length were 662.38±2.54 g and 71.48±0.12 mm, respectively. Sex had significant influence on live weight, thigh and wing proportions wherein cocks had heavier and stronger legs and wings. Abdominal fat was significantly higher in females. The egg production up to 40 and 52 weeks of age was 80.29±0.03 and 133.16±0.05 eggs, respectively. The egg mass up to 52 weeks of age was 7447.41±2.98 g in PD-2 line. The heritability was low up to 15% for egg production and 17% for egg mass, the primary trait of selection in PD-2 line. Egg mass and egg weights at different ages had positive association as larger and high number of eggs contributed more to the egg mass. The egg mass and egg production had a significant positive association in PD-2 line. The egg quality was reasonably fair with average shape index of 75.22 and Haugh unit of 79.0±0.62. The albumen, yolk and shell weights were 30.20±0.14, 15.8±0.09, 0.40±0.02 g, respectively. The proportion of heritability and the magnitude of association between the important traits provide requisite information for implementing the breeding program for improvement of egg production in PD-2 line.

Growth Patterns, Body Weight, and Morphometric of KUB Chicken, Sentul Chicken and Arab Chicken

The purpose of this research was to determine the body weight, weight gain and morphometric characteristics of several local chicken strains. The research materials were KUB chicken, Sentul chicken, and Arab chicken. The method used was an experiment with a sample of 82 chickens from each strain. Data collected include body weight, weight gain, beak length, beak width, head length, head circumference, head height, neck length, neck circumference, wing length, back length, back height, chest length, chest width, shank length, shank circumference, tibia length, tibia circumference, third finger length, and pubic bone distance. Data collected were analyzed using the average difference test (t-test). The average value vector of body measurements of chicken lines was analyzed using the T²-Hotelling statistical test. Main Component Analysis statistical test was used to identify the shape and size characteristics of each chicken strain. The results showed that the body weight and morphometrics of KUB chickens at 3 months were significantly different (P&lt;0.05) with Sentul chickens and Arab chickens. KUB chicken`s body weight gain at 3 months was not significantly different (P&gt;0.05) from Sentul chickens, but it was significantly different (P&lt;0.05) from Arab chickens. Body weight, weight gain, and morphometrics of KUB chickens were higher than Sentul chickens and Arab chickens. The size indicator in KUB chickens was the tibia length while in Sentul chickens and Arab chickens was chest length. The characteristic of KUB chickens was the back length, Sentul chickens was tibia length, and Arab chickens was the shank circumference.

Identification of single nucleotide polymorphism in heat shock protein HSP70 and HSP90 after four selection generations in two lines of chickens

Climatic changes have affected the entire world resulting in high temperatures, consequently leading to a negative impact on poultry production. Therefore, we conducted this study to detect the single nucleotide polymorphisms (SNPs), which is responsible for producing heat shock protein HSP70 and HSP90 in two selected lines of chicken, Dandarawi and Leghorn that were formally selected for four generations, for achieving a better growth rate under heat pressure. At age 16 weeks, body weight and body measurements were evaluated for Dandarawi and Leghorn males. mRNA levels of HSP70 and HSP90 genes were measured, and the polymorphism of these two genes was examined via DNA sequencing. Results showed that after four generations under heat stress conditions, the Dandarawi and Leghorn lines of chicken were distinguishable in terms of body weight and body measurements. Additionally, HSP70 and HSP90 a highly significant increase in gene expression for the Dandarawi line compared to the Leghorn line. These results indicate that the Dandarawi breed has genes that enable it to adapt under hot climate conditions. Hence, when selected to increase production under high temperatures, the gene expression is twice that of the Leghorn strain under the same conditions of selection. The Dandarawi chicken had better heat tolerance than the Leghorn chicken. The difference was observed on chromosome 5 in the region 52784621: 52784671 in exon 1 in HSP70 that was caused by the amino acid cysteine instead of arginine because of the substitution of T nucleotide with C, indicating the alteration in the gene expression level of HSPs. Therefore, we recommend using chromosome 5 in the region 52784621: 52784671 in exon 1 in HSP70 as a candidate gene in selection programs for evaluating heat stress tolerance.

Genetic evaluation of growth and production performance and short term selection response for egg mass in Gramapriya female line chicken

A comprehensive study was carried out in Gramapriya female line (PD-3) with respect to growth, production, selection response and population structure utilizing the data generated for the last 7 generations at ICAR-Directorate of Poultry Research, Hyderabad, Telangana. The average fertility was 71.89% and hatchability was 80.52% on fertile egg set and 57.08% on total egg set. The least squares means (LSMs) for body weight at 4 and 6 weeks of age were 168.06±0.01 and 320.10±0.02 g, respectively. The heritability estimates from sire and dam components of variance were moderate to high for both body weight and shank length (0.27 to 0.35). The correlation coefficients (genetic and phenotypic) between body weights and shank length were positive and high in magnitude except with day old body weight. Age at sexual maturity (ASM) was 171.47±0.01 days in PD-3 population. The part period egg production (EP 40) at 40 weeks of age was 75.60±0.01 eggs with an egg mass (EM 40) of 4,157.19±0.99 g. The h2 estimates for ASM, EW 40, EP 40 and EM 40 were 0.16±0.07, 0.42±0.11, 0.18±0.07 and 0.15±0.06, respectively, which were moderate to high in magnitude. The egg production at 40 weeks of age was negatively associated with ASM, body weight and egg weight at different ages. Egg mass had significant positive association with egg production and egg weight at 40 weeks of age. The selection intensity (i) ranged from 0.32 to 0.85 while, rate of inbreeding ranged between 0.0031 and 0.0033 in PD-3 line. The direct selection response was 151.54 g on genetic scale and 79.75 on phenotypic scale for primary trait of selection, EM 40 over the last 7 generations. Realized and estimated heritability estimates were 0.29 and 0.15, respectively for EM 40. The study concluded that, PD-3 population was in ideal condition with respect to growth and production performance. Positive selection response for primary trait EM 40 was observed in the population maintaining both egg production and egg weight, the important traits for sustainable rural poultry farming.

Genome-wide characterization of copy number variations in the host genome in genetic resistance to Marek\u2019s disease using next generation sequencing

BackgroundMarek’s disease (MD) is a highly neoplastic disease primarily affecting chickens, and remains as a chronic infectious disease that threatens the poultry industry. Copy number variation (CNV) has been examined in many species and is recognized as a major source of genetic variation that directly contributes to phenotypic variation such as resistance to infectious diseases. Two highly inbred chicken lines, 63 (MD-resistant) and 72 (MD-susceptible), as well as their F1 generation and six recombinant congenic strains (RCSs) with varied susceptibility to MD, are considered as ideal models to identify the complex mechanisms of genetic and molecular resistance to MD.ResultsIn the present study, to unravel the potential genetic mechanisms underlying resistance to MD, we performed a genome-wide CNV detection using next generation sequencing on the inbred chicken lines with the assistance of CNVnator. As a result, a total of 1649 CNV regions (CNVRs) were successfully identified after merging all the nine datasets, of which 90 CNVRs were overlapped across all the chicken lines. Within these shared regions, 1360 harbored genes were identified. In addition, 55 and 44 CNVRs with 62 and 57 harbored genes were specifically identified in line 63 and 72, respectively. Bioinformatics analysis showed that the nearby genes were significantly enriched in 36 GO terms and 6 KEGG pathways including JAK/STAT signaling pathway. Ten CNVRs (nine deletions and one duplication) involved in 10 disease-related genes were selected for validation by using quantitative real-time PCR (qPCR), all of which were successfully confirmed. Finally, qPCR was also used to validate two deletion events in line 72 that were definitely normal in line 63. One high-confidence gene, IRF2 was identified as the most promising candidate gene underlying resistance and susceptibility to MD in view of its function and overlaps with data from previous study.ConclusionsOur findings provide valuable insights for understanding the genetic mechanism of resistance to MD and the identified gene and pathway could be considered as the subject of further functional characterization.

Comparing the innate and humoral immune responses of different chicken lines to Infectious Bronchitis Virus (IBV) infection

Infectious Bronchitis Virus (IBV) is a gammacoronavirus that is prevalent in commercial chicken flocks, resulting in characteristic clinical signs including snicking, rales, decreased tracheal ciliary activity, reduced weight gain and reduced egg production. Preliminary results indicate that there is a different clinical response to IBV infection in different chicken lines. Therefore, we aim to determine whether there is a differential innate or humoral immune response to IBV between chicken lines. A series of in vivo experiments were conducted comparing brown leghorns (Rhode Island Red, RIR (Roslin)) to white leghorns (from Valo and Ovagen). Trachea and bursa were collected from infected and control birds at four-, six- and fourteen-days post infection (dpi). There was a difference in snick rate and rales between the RIRs and the white leghorns (both lines). However, no difference was observed in ciliary activity. Viral load was determined by absolute quantification using qRT-PCR. The viral load in the trachea of RIRs was significantly lower (p&lt;0.05) at 6 dpi compared to 4 dpi, unlike in Ovagen birds where there was no significant difference between the timepoints. Relative gene expression of IFN-α, IFN-β, IFN-γ, IL-6 and IL-1β in these tissues will be measuredby qRT-PCR. Serum was processed from whole blood collected at zero and ten dpi for use in IBV specific ELISAs which will measure antibody responses in the chicken lines. This project aims to explore immune responses against IBV as well as identifying the causes of variability in experimentation using chickens to investigate IBV infection.

Fatty Acid Composition and Regulatory Gene Expression in Late-Term Embryos of ACRB and COBB Broilers.

Cobb broilers (COBB) have been heavily selected for their production performance in the past several decades, while the Athens Canadian Random Bred (ACRB) chickens, a meat-type breed, have been kept as a non-selected control strain. The purpose of this study was to compare these two lines of chickens at late embryonic development and identify the molecular markers and fatty acid profiles underlining their differences in growth performance due to selection. Fertilized eggs of the ACRB (n = 6) and COBB (n = 6) were used at 14 and 18 embryonic days. Genes involved in lipogenesis and myogenesis were measured using quantitative real-time reverse transcroption-polymerase chain reaction (RT-PCR), and fatty acid (FA) compositions of egg yolk, muscle, and liver were measured using gas chromatography. COBB had higher egg weight, embryo weight, and breast and fat ratio. The gene expression in the liver showed an interaction between age and breed on FASN expression, with the highest level in COBB at E18. ACRB had higher ApoB and MTTP expression, but lower SREBP-1 expression compared to COBB. No difference was found in myogenesis gene expression in the muscle between two breeds. For the FA composition, muscle was largely affected by both breed and age. Yolk and liver were affected mainly by breed and age, respectively. Constant interaction effects in docosahexaenoic acid (DHA), indicating the highest level in all the tested tissues of ACRB at E14 and the constant main effects with higher myristic, palmitic, and gondoic, but lower linolenic acid in the liver and yolk of COBB compared to the levels in those of ACRB. Finally, fat accumulation in the liver had no obvious difference between the breeds but was higher when embryo was older. In conclusion, broiler breed affects egg, embryo, and tissue weight, as well as FA composition in initial egg yolk and throughout the embryonic development. The highest docosahexaenoic percentage was observed in ACRB, indicating that genetic selection may result in fatty acid profile changes such as lower DHA content in chicken tissues and eggs.

Major histocompatibility complex B variability in Korean native chicken breeds

Adaptive genetic variations have direct influences on the fitness traits of the animal. The major histocompatibility complex B (MHC-B) region is responsible for adaptive and innate immune responses in chickens. In native Korean chicken breeds, no information on serologically defined B haplotypes is available. We investigated the MHC-B diversity in 5 restored lines of Korean native chicken and Ogye chicken breeds using a recently described MHC-B single-nucleotide polymorphism (SNP) panel and the MHC-linked LEI0258 variable number of tandem repeat marker. High SNP haplotype diversity was observed in Korean native chicken breeds with an average of 9.7 MHC-B SNP haplotypes per line. The total number of haplotypes ranged from 6 to 12 per line, and population-specific haplotypes ranged from 3 to 4. A total of 41 BSNP haplotypes, including 26 novel population-specific haplotypes and 15 common haplotypes, were reported over all populations. The 15 common haplotypes included 7 novel and 8 previously reported standard haplotypes. Selection and breeding evidence supports the observation of common haplotypes between the Korean native chicken and exotic breeds. Similarly, the LEI0258 marker showed allele variation, between 193 bp and 474 bp having 5 to 8 alleles per population. Some of these alleles (193, 249, 309, and 443 bp) were shared and more frequently observed. Comparison between SNP haplotypes and LEI0258 allele sizes for the same samples showed that some LEI0258 allele sizes correspond to more than one BSNP haplotype. The use of the MHC-B SNP panel greatly enhances the identification of MHC diversity compared with the sole use of the LEI0258 marker in native chicken populations.

Current state and problems of poultry breeding in Russia (review)

Modern poultry breeding in Russia is one of the fastest growing sectors of agriculture, but the prosperity of the industry is almost entirely dependent on supplies of breeding material from abroad. Russia practically has no its own breeding base in both egg- and meat-type commercial crosses of chickens. Most of the domestic commercial crosses that had occupied leading positions in Russian poultry breeding have been lost now. More than 90 % of commercial lines of breeding stocks in Russia are imported. Foreign poultry breeding companies merge into transnational holdings engaged in multi-species breeding of farm animals, which allows them not to depend on market conditions in the industry. The reverse side of such a consolidation on a global scale is a decrease in the genetic diversity of poultry and a high level of inbreeding in commercial chicken lines. In these circumstances, there is a real biological danger for the preservation of these lines due to the potential susceptibility of “monocultures” to new diseases, which can eliminate the genetically homogeneous population. Any selection system is based on an assessment of the breeding value of potential parents. Its purpose is to obtain, as far as possible, the most accurate forecast of the genetic value of an individual and the productive qualities of its progeny. These requirements are optimally met by the BLUP methodology, in which molecular genetics (SNP) data can be successfully integrated, which allows supplementing the statistical analysis with genomic selection technologies. This is especially true for traits that cannot be measured, or can only be measured in one sex, or only at the end of the productive period. The inclusion of genomic selection methods in breeding programs makes it possible significantly increase the selection efficiency for the main economical traits of chickens. The main task in the Russian breeding poultry industry is the creation of its own competitive breeding base, not inferior to Western commercial crosses in terms of productivity. To create poultry breeding centers and grandparents/parents-breeding farms the state support is needed. It is also necessary to develop and implement innovative methods in the field of genomic selection, as well as software and information systems and specialized selection computer programs for processing and analyzing meta-data.

Production of Germ-Free Fast-Growing Broilers from a Commercial Line for Microbiota Studies.

Studies of the gut microbiota contribution to the host physiology and immunocompetence are facilitated by the availability of germ-free animal models, which are considered the gold standard. Nesting birds are ideal models for the production of germ-free animals since there is no need to raise their relatives under sterile conditions. Germ-free chickens are mainly generated from specific-pathogen-free (SPF) experimental lines, which are poorly representative of commercial chicken lines.The method proposed here allowed the production of germ-free chickens from the fast growing broiler line Ross PM3, commonly used by the poultry industry.Eggs were quickly collected after laying at a broiler breeder farm.They underwent a strict decontamination process from the collection to the introduction in a sterile egg hatching isolator.The chicks have been hatched and kept in these sterile isolators during the period necessary to control their sterility. Originally developed for an experimental SPF white leghorn line, the present protocol has been adapted not only to the Ross PM3 broiler line but also to quails. It therefore represents a robust and readily adaptable procedure to other poultry species and nesting birds of economic, biological or ecological relevance.